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Polyphenolic additives in sequencing by synthesis

  • US 10,337,050 B2
  • Filed: 02/08/2017
  • Issued: 07/02/2019
  • Est. Priority Date: 02/11/2016
  • Status: Active Grant
First Claim
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1. A method of incorporating labeled nucleotides, comprising:

  • a) providingi) a plurality of nucleic acid primers and template molecules,ii) a polymerase,iii) a cleave reagent comprising a reducing agent and the polyphenolic compound gallic acid, andiv) a plurality of nucleotide analogues wherein at least a portion of said nucleotide analogues is labeled with a label attached through a cleavable disulfide linker to the base;

    b) hybridizing at least a portion of said primers to at least a portion of said template molecules so as to create hybridized primers;

    c) incorporating a first labeled nucleotide analogue with said polymerase into at least a portion of said hybridized primers so as to create extended primers comprising an incorporated labeled nucleotide analogue;

    d) detecting said incorporated labeled nucleotide analogue; and

    e) cleaving the cleavable linker of said incorporated nucleotide analogues with said cleave reagent.

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